高产蛋白酶和淀粉酶米曲霉菌株的选育

从J2、M2和Q2三株米曲霉菌中选择出总酶活力最高的J2作为出发菌株,通过紫外诱变后,得到突变菌株Y3,其酸性蛋白酶活力由出发菌株的313 U/g提高至986 U/g,提高了215％,碱性蛋白酶活力由出发菌株的6936 U/g提高至10385 U/g,提高了49.7％,中性蛋白酶活力由出发菌株的5675 U/g提高至9034 U/g,提高了59.2％,且淀粉酶活力也由出发菌株的284 U/g提高至412U/g,提高了45.1％.经传代实验表明,突变菌株Y3的遗传特性稳定.     

Response surface optimization and characteristics of Indica rice starch‐based fat substitute prepared by α‐amylase

Response surface methodology (RSM) was used to study the effect of enzyme to substrate ratio (11.8–23.6 U α‐amylase/g rice starch), hydrolysis temperature (90–100°C) and pH value (6.0–6.6) on the gel strength of rice starches‐based fat substitute using α‐amylase hydrolysis. The optimum conditions obtained from response surface analysis was 16.52 U/g enzyme dosage, 92°C hydrolysis temperature while the influence of pH was found insignificant in the range tested. Under these optimum conditions, the gel strength of this fat substitute was 113 g/cm², very close to the gel strength of butter of 114 g/cm², while the solubility of the substitute was 1.33 ± 0.01% and the swelling power 4.85 ± 0.02. There were no observable differences in the granular size distribution between the untreated rice starch and the hydrolyzed rice starch. Rheological properties of this rice starch‐based fat substitute implied that it is easier for the substitute to form three‐dimensional networks under 34°C.     

Changes in selected physical property and enzyme activity of rice and barley koji during fermentation and storage

Koji are solid-state fermentation products made by inoculating steamed grains with the spores of fungi, particularly Aspergillus spp. This research was undertaken to identify the fermentation and storage conditions optimal for the production and maintenance of selected hydrolytic enzymes, such as α-amlyase and protease, in koji. Steamed rice and barley were inoculated with 2 × 10 (11) Aspergillus oryzae spores per kilogram of grains and fermented for 118 h in a growth chamber at 28 to 32 °C with controlled relative humidities. Samples were drawn periodically during fermentation and storage at -20, 4, or 32 °C, and α-amylase and protease activity, mold counts, a(w), moisture contents, and pH of collected samples were determined. It was observed that the a(w), moisture contents, and pH of the koji were influenced by the duration of fermentation and temperature of storage. The α-amylase activity of both koji increased as the populations of A. oryzae increased during the exponential growth phase. The enzyme activity of barley koji was significantly higher than that of rice koji, reaching a peak activity of 211.87 or 116.57 U at 46 and 58 h, respectively, into the fermentation process. The enzyme activity in both products started to decrease once the mold culture entered the stationary growth phase. The protease activities of both koji were low and remained relatively stable during fermentation and storage. These results suggest that rice and barley koji can be used as sources of α-amylase and desired enzyme activity can be achieved by controlling the fermentation and storage conditions. PRACTICAL APPLICATION: Amylases and proteases are 2 important hydrolytic enzymes. In the food industry, these enzymes are used to break down starches and proteins while reducing the viscosity of foods. Although amylases and proteases are found in plants and animals, commercial enzymes are often produced using bacteria or molds through solid state fermentation, which is designed to use natural microbial process to produce enzymes in a controlled environment. A properly produced and maintained koji with a high hydrolytic enzyme activity can serve as an important source of the enzymes for the food industry.     

Physicochemical characterisation and α‐amylase inhibitory activity of tea polysaccharides under simulated salivary,gastric and intestinal conditions

Tea polysaccharides (TPS) are one of the main components of tea with various bioactivities. Digestion could result in the changes of the physicochemical properties and bioactivities of polysaccharides. The objective of this study was to determine the changes in physicochemical properties and α‐amylase inhibitory activities of TPS after simulated digestion. The structure of TPS determined by UV, IR and SEM showed that no obvious changes after salivary digestion, but significant changes after gastrointestinal digestion were observed. After intestinal digestion, the contents of reducing sugar were increased from 0.650 ± 0.011 to 1.594 ± 0.078 mm , and the centre molecular weight was decreased from 540.57 to 375.35 KDa. The molar ratio of monosaccharide was changed after digestion. And α‐amylase inhibition rate was significantly increased (P < 0.05). The results could provide information on the digestibility of TPSin vitro and contribute to the development of related functional foods or medicines.     

Porous‐structured extruded instant noodles induced by the medium temperature α‐amylase and its effect on selected cooking properties and sensory characteristics

To enhance the rehydration and palatability characteristics of extruded noodles, a new type of instant noodles with a well‐developed porous structure was successfully created by cooperating medium temperature α‐amylase (MTA) with extrusion technique. To explain the formation reason of the porous structure, a detailed mechanism analysis was firstly carried out. It turned out that a portion of starch was degraded into water‐soluble dextrin during the enzymatic extrusion treatment. Besides, a slight restriction of the excessive MTA on the starch gelatinisation was demonstrated. The appearance of the well‐developed porous structure was attributed to the leaching of dextrin and loosely bound starch fragments. Therefore, the extruded noodles with 0.40‰ MTA concentration were recommended based on the results of cooking quality, textural characteristics and sensory evaluation.     

Interaction of aqueous extracts of two varieties of Yam tubers (Dioscorea spp) on some key enzymes linked to type 2 Diabetes in vitro

This study sought to assess the effect of processing of yam flour [from white yam (Dioscorea rotundata) and water yam (Dioscorea alata)] to paste, on the antioxidant properties and inhibition of key enzymes linked to type‐2 diabetes (α‐amylase and α‐glucosidase). The phenolic content (phenol and flavonoid), antioxidant properties, α‐amylase and α‐glucosidase inhibitory properties of the aqueous extracts of the yam products were determined. Processing into paste caused significant (P < 0.05) percentage decrease in their total phenol and flavonoid content of the yam varieties. However, significant increase (P < 0.05) in the antioxidant properties of the yam varieties was observed after processing. Furthermore, the yam extracts inhibited α‐amylase and α‐glucosidase activities in vitro in a dose‐dependent parttern (1–4 mg mL^?1), however, the pasting process caused significant (P < 0.05) decrease in the α‐amylase inhibitory and α‐glucosidase inhibitory activities. Therefore, processing the yam varieties into paste (browned) could potentially increase their antioxidant capacity as well as decreasing their α‐amylase and α‐glucosidase inhibitory properties.     

Inhibitory potential of phenylpropanoid glycosides from Ligustrum purpurascens Kudingcha against α‐glucosidase and α‐amylase in vitro

The leaves of Ligustrum purpurascens are used in a Chinese traditional tea called small‐leaved kudingcha, which is rich in phenylpropanoid glycosides (PPGs) and has many beneficial properties. Two critical exoacting glycoside hydrolase enzymes (glucosidases) involved in carbohydrate digestion are α‐glucosidase and α‐amylase. We investigated the properties of PPGs from L. purpurascens for inhibiting α‐amylase and α‐glucosidase activity in vitro and found IC₅₀ values of 1.02 and 0.73 mg mL^?1, respectively. The patterns of inhibiting both α‐amylase and α‐glucosidase were mixed‐inhibition type. Multispectroscopy and molecular docking studies indicated that the interaction between PPGs and α‐amylase and α‐glucosidase altered the conformation of enzymes, with binding at the site close to the active site of enzymes resulting in changed enzyme activity. Our studies may help in the further health use of small‐leaved kudingcha.     

Alpha-amylase inhibitor from local common bean selection: Effect on growth and development of Corcyra cephalonica

Cultivars of common bean (Phaseolus vulgaris L.) from the Kalpa valley of the Kinnaur district were screened for alpha-amylase inhibitor activity against porcine pancreatic α-amylase and significant differences were found amongst them. The inhibitor protein was partially purified from Triloki cultivar by ammonium sulphate precipitation and Sephadex G-100 gel filtration chromatography. Inhibitor protein was non-competitive, heat labile and retained 55.51 per cent activity at 70 °C. It had two pH optima of 5.0 and 7.6. Inhibitor protein inhibited amylases of 3rd -4th instar larval extracts of Corcyra cephalonica Stainton, the rice moth. C. cephalonica larvae (3rd -4th instar) were fed on wheat flour mixed with 0.013 percent (w/w) of partially purified α-amylase inhibitor protein. The growth of larvae in the treatment was significantly reduced. Treated larvae showed 100 percent mortality after 11 days. In the untreated control, no larval mortality was observed and adults emerged after 45 days. The inhibitor protein was effective at very low concentrations and could be exploited for control of this grain storage pest.     

Influence of a reduced-starch diet with or without exogenous amylase on lactation performance by dairy cows

The objective of this trial was to determine lactation performance responses in high-producing dairy cows to a reduced-starch versus a normal-starch diet and to the addition of exogenous amylase to the reduced-starch diet. Forty-five multiparous Holstein cows, 68 ± 29 d in milk and 696 ± 62 kg of body weight (BW) at trial initiation, were randomly assigned to 1 of 3 treatments in a completely randomized design; a 2-wk covariate adjustment period with cows fed the normal-starch diet was followed by a 10-wk treatment period with cows fed their assigned treatment diets. The normal-starch total mixed ration did not contain exogenous amylase (NS−). The reduced-starch diets, formulated by partially replacing corn grain and soybean meal with whole cottonseed and wheat middlings, were fed without (RS−) and with (RS+) exogenous amylase addition to the total mixed ration. All diets contained 50% forage and 19.8% forage neutral detergent fiber (dry matter basis). Starch and neutral detergent fiber concentrations averaged 27.0 and 30.9%, 22.1 and 35.0%, and 21.2 and 35.3% (dry matter basis) for the NS−, RS−, and RS+ diets, respectively. Expressed as a percentage of BW, dry matter intake was greater for cows fed RS− than for cows fed NS− or RS+. Intake of neutral detergent fiber ranged from 1.09 to 1.30% of BW among the treatments, with that of RS− being 21% greater than that of NS−. Milk yield tended to be greater for cows fed NS− compared with the RS diets. Milk fat content and yield were unaffected by treatment. Milk protein content and yield were greater for cows fed NS− compared with the RS diets. Concentrations of milk urea nitrogen were greater for cows fed RS diets compared with the NS− diet. Body weight, BW change, and body condition score were unaffected by treatment. Feed conversion (kg of milk/kg of dry matter intake) was 10% greater on average for cows fed NS− than for cows fed the RS diets, and tended to be 6% greater for cows fed RS+ compared with RS−. Feeding a reduced-starch diet formulated by partially replacing corn grain and soybean meal with a wheat middlings and whole cottonseed mixture compared with a normal-starch diet without addition of exogenous amylase to either diet reduced milk and component-corrected feed conversions. Addition of exogenous amylase to a reduced-starch diet was of minimal benefit in this study.